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Journal of Virology, March 2004, p. 2601-2605, Vol. 78, No. 5
0022-538X/04/$08.00+0     DOI: 10.1128/JVI.78.5.2601-2605.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Human Immunodeficiency Virus Type 1 Escapes from RNA Interference-Mediated Inhibition

Atze T. Das,^1, Thijn R. Brummelkamp,^2, Ellen M. Westerhout,¹ Monique Vink,¹ Mandy Madiredjo,² René Bernards,² and Ben Berkhout^1*

Department of Human Retrovirology, Academic Medical Center, University of Amsterdam,¹ Division of Molecular Carcinogenesis, The Netherlands Cancer Institute, Amsterdam, The Netherlands²

Received 3 September 2003/ Accepted 14 October 2003

Short-term assays have suggested that RNA interference (RNAi) may be a powerful new method for intracellular immunization against human immunodeficiency virus type 1 (HIV-1) infection. However, RNAi has not yet been shown to protect cells against HIV-1 in long-term virus replication assays. We stably introduced vectors expressing small interfering RNAs (siRNAs) directed against the HIV-1 genome into human T cells by retroviral transduction. We report here that an siRNA directed against the viral Nef gene (siRNA-Nef) confers resistance to HIV-1 replication. This block in replication is not absolute, and HIV-1 escape variants that were no longer inhibited by siRNA-Nef appeared after several weeks of culture. These RNAi-resistant viruses contained nucleotide substitutions or deletions in the Nef gene that modified or deleted the siRNA-Nef target sequence. These results demonstrate that efficient inhibition of HIV-1 replication through RNAi is possible in stably transduced cells. Therefore, RNAi could become a realistic gene therapy approach with which to overcome the devastating effect of HIV-1 on the immune system. However, as is known for antiviral drug therapy against HIV-1, antiviral approaches involving RNAi should be used in a combined fashion to prevent the emergence of resistant viruses.

A.T.D. and T.R.B. contributed equally to this work.